Our objective is to characterize the adenylate cyclase of Bordetella pertussis, to clone its gene, and to use the gene to produce more of the protein than can be recovered from the bacterium. We have found that this enzyme is more complex than previously thought. Our biochemical studies allowed us to develop affinity procedures which we will use to purify this enzyme. Our initial experiments have also determined that binding of calmodulin is direct and we have developed methods to screen for the gene in recombinant DNA libraries based on the ability of the adenylate cyclase peptide to bind calmodulin. Cloned sequences will be characterized by sequencing and then manipulated for several purposes which include the production of enzyme in E. coli, the creation of specific Bordetella adenylate cyclase mutants, and the expression of the gene in eucaryotic cells. After physical characterization to resolve some of the conflicting results in the literature, we will study action of the adenylate cyclcase on mammalian target cells. Finally we will take advantage of the unusual evolution of the gene and attempt to use it for long term induction of adenylate cyclase and cAMP in mammalian cells.